CN101530467A - Protection effect of compound Danshen dropping pill on cordis microcirculatory disturbance and myocardial damage - Google Patents
Protection effect of compound Danshen dropping pill on cordis microcirculatory disturbance and myocardial damage Download PDFInfo
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- CN101530467A CN101530467A CN200810052432A CN200810052432A CN101530467A CN 101530467 A CN101530467 A CN 101530467A CN 200810052432 A CN200810052432 A CN 200810052432A CN 200810052432 A CN200810052432 A CN 200810052432A CN 101530467 A CN101530467 A CN 101530467A
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Abstract
The invention relates to the new application of Chinese traditional medicine products, in particular to the application of compound Danshen dropping pill in the treatment and prevention of cordis microcirculatory disturbance and myocardial damage. The test data indicate that the one-time pre-addition of the compound Danshen dropping pill can improve the microcirculatory disturbance caused by ischemia-reperfusion and the myocardial damage, and repeated dosing of the compound Danshen dropping pill can prevent the microcirculatory disturbance caused by ischemia-reperfusion and the myocardial damage.
Description
Technical field:
The present invention relates to the new purposes of tcm product, the particularly application of FUFANG DANSHEN DIWAN in treating and/or preventing cardiac microcirculation disturbance and myocardial damage.
Background technology:
Along with the raising of people's living standard and the change of living habit, cardiovascular disease has become the main diseases kind that threatens human health and life, influence medical treatment finance.The annual whole world is because of dead nearly 1,650 ten thousand people of number of cardiovascular diseases, and wherein, about 3,000,000 people of China account for 45% of total death toll.
Department of cardiac surgery extracorporeal circulation, bypass operation of coronary artery, complicated congenital heart disease are just controlled the clinical practice of interventional therapies such as art, prosthetic valve replacement and trunk surgical operation, saved cardiovascular patient's life in a way, but, (ischemia/reperfusion, I/R) damage then becomes a main difficult problem that hinders cardiovascular surgical operation late result to the postoperative ischemia-reperfusion.On the other hand, the ischemical reperfusion injury of the link of recovery after the sudden cardiac arrest and cardiovascular spasm initiation has also influenced rescued effect.Therefore preventing and improving myocardial ischemia reperfusion injury is to improve the important step that cardiovascular disease is just controlled effect, reduced its mortality rate.
When heart coronaries blood blocks because of factors such as spasm, arteriosclerosis and thrombosis, caused and blocked dirty ischemia and anoxia.ATP generates and reduces during ischemia, and the generation hypoxanthine is decomposed in the AMP metabolism, because intracellular calcium increases, the activator protein enzyme makes xanthine dehydrogenase irreversibly be transformed into xanthine oxidase.Behind thrombolytic, vasodilation or interventional therapy, blood flow logical again (perfusion again) provides O
2The time, hypoxanthine and O
2Reach water under xanthine oxidase catalysis, a large amount of negative oxygen anions of generation (
O
2 -),
O
2 -Under the effect of superoxide dismutase (SOD), change into hydrogen peroxide (H
2O
2), the latter changes into H under the effect of catalase (CAT)
2O, a part
O
2 -And H
2O
2Through Haber-Weiss reaction change into hydroxy radical (
OH).In addition, the NO that produces of blood vessel endothelium with
O2
-Be combined into ONOO
-H
2O
2,
OH, ONOO
-Deng being the very strong peroxide of toxicity all, can cause lipid peroxidation, dna break etc. to cause blood vessel endothelium and myocardial cell injury.The peroxide that I/R produces, by degraded transcription factor Profilin-κ B (I-κ B), active nuclei is transcribed transcription factor (NF-κ B), causes its subunit P65, and the consideration convey of P50 moves, and particularly the consideration convey of P50 moves to start and transfers the synthetic of the associated protein of dying.The expression of blood vessel endothelium choosing folding plain (E-selectin) and leukocyte choosing folding plain (L-selectin) causes that leukocyte rolls along blood vessel wall, and the expression of blood vessel endothelium adhesion molecule ICMA-1 and LAM CD11b/CD18 causes sticking of leukocyte and blood vessel wall.Stick with blood vessel wall on leukocyte further produce peroxide by nadph oxidase on the one hand, simultaneously, again can extracellular proteinase, increase the weight of the damage of cardiovascular and myocardial cell.The leukocyte that attaches on the blood vessel wall is swum out of outside blood vessel by the blood vessel endothelium of vascular endothelial cell gap or damage, increases the weight of the damage of perivascular cells.The damage of vascular endothelial cell and basement membrane of blood vessel has increased vascular permeability, causes leaking outside of plasma albumin, cause around the blood vessel between the edema of matter.The consideration convey of P50 moves the synthetic of inflammatory mediators such as all right induced tumor necrosis factor-alpha (TNF-α), IL-6 and discharges, these inflammatory factors are further induced the proteic startup of dying of the expression of adhesion molecule and accent, increase the weight of cardiac microcirculation disturbance and myocardial cell injury.Cardiac microcirculation disturbance that ischemia-reperfusion causes and myocardial cell injury are the pathological change process of the too many levels of complexity, the interaction of oxygen-derived free radicals, calcium overload, leukocyte and vascular endothelial cell, inflammatory mediator, apoptosis protein regulation and control etc. all participate in this process, and peroxide degradation I-κ B, activation NF-κ B, cause its subunit P65, or the consideration convey of P50 moves, and particularly to move be to increase the weight of to pour into the back microcirculation disturbance again and myocardial cell is transferred one of key link of dying to the consideration convey of P50.
Had at present about adenosine, calcitonin-gene-related peptide, Kallidin I, PGI2, NO etc. for the prevention of heart I/R damage and the research of improvement effect; but; because it only can act on some links or target spot in the complicated heart I/R damage process, the effect after improving heart I/R aspect heart microcirculation disturbance and the myocardial cell protection is still unsatisfactory.
FUFANG DANSHEN DIWAN (CP) is made up of the active ingredient of Radix Salviae Miltiorrhizae, Radix Notoginseng, Borneolum Syntheticum, is mainly used to treat cardiovascular disease in states such as China, Korea S, Russia and Vietnam.Radix Salviae Miltiorrhizae is the dry root of Labiatae clary Salviamiltiorrhiza, and the main component of Radix Salviae Miltiorrhizae aqueous solution comprises danshensu, protocatechualdehyde, protocatechuic acid, caffeic acid, salvianolic acid A-G and alkannic acid.Wherein, danshensu is its basic structure, has participated in the structure of salvianolic acid A-G and alkannic acid structure.Former studies shows, after the filling stomach gives the Radix Salviae Miltiorrhizae total acid, can detect danshensu and caffeic acid in the serum.Show that with in vitro study Radix Salviae Miltiorrhizae passes through microcirculation improvement in the various bodies, coronary artery dilator improves blood flow, and the protection cardiac muscle prevents myocardial ischemia reperfusion injury.Its mechanism is mainly activation and the overload that Radix Salviae Miltiorrhizae can suppress calcium ion in the myocardial cell, removes oxygen-derived free radicals, suppresses apoptosis of cardiac muscle and protection vascular endothelial cell etc.The water-soluble components of Radix Salviae Miltiorrhizae can improve Rats survival rate behind the I/R; and reduce the left ventricle infarct size; alkannic acid B can reduce the degree of myocardial damage behind the rabbit heart ischemia, and salvianolic acid A and salvianolic acid B can be protected cerebral lesion, and FUFANG DANSHEN DIWAN can be improved the hepar damnification that I/R causes.Danshensu can also reduce mitochondrial membrane damage and the lipid peroxidation that is caused by I/R, the superoxide anion that removing is produced by xanthine and xanthine oxidase (
O2-), salvianolic acid A can suppress by the inductive brain LPO of I/R and remove hydrogen peroxide free radical (OH
-), salvianolic acid B can be removed 1.1-diphenyl-2-picrylhydrazyl, suppress the peroxide in LPO and removing rat hepatocytes and the stellate cells, be suppressed at the generation of the peroxide that causes by amyloid peptide A in the PCL2 cell, suppress the inductive human aortic smooth muscle cell's peroxide of TNF-α and produce and the nadph oxidase activity, alkannic acid magnesium B can remove ONOO-.In addition, the consideration convey that the water soluble ingredient of Radix Salviae Miltiorrhizae can also suppress the inductive NF-κ of TNF-α B moves, protocatechualdehyde, salvianolic acid B can suppress the expression of the mRNA of the expression of the ICAM-1 of the inductive HUVEC of TNF-α and VCAM-1 and ICAM-1 and VCAM-1, suppress the dna binding activity that TNF-α induces NF-κ B and AP-1.Radix Salviae Miltiorrhizae, salvianolic acid B and Radix Salviae Miltiorrhizae aqueous solution can suppress to be caused by TNF-α the expression of the E-selection of vascular endothelial cell, suppressed to CP dosage interdependence the expression of ICAM-1 and VCAM-1 among the HUVEC that TNF-α causes, the synthetic and cell proliferation of DNA in vascular smooth muscle cell that suppresses simultaneously that growth factor B B that platelet produces causes.Radix Salviae Miltiorrhizae can also suppress the activation and the overload of calcium ion in the myocardial cell, suppresses apoptosis of cardiac muscle.
Radix Notoginseng is the dry root of Panax notoginseng, Radix Notoginseng total arasaponins (PanaxnotogisengSaponins PNS) is main active component in the Radix Notoginseng, mainly comprise 30 surplus kind of saponin component.Be widely used in the treatment of cardiovascular and cerebrovascular disease, hepatic insufficiency etc. and microcirculation disturbance relevant disease clinically.Its mechanism may with the immunoloregulation function of Radix Notoginseng, the microcirculation improvement obstacle suppresses the expression of hematoblastic gathering and adhesion factor, reduces leukocyte adhesion in blood vessel endothelium and to improve the function of endothelium relevant.The Chinese medicine compound preparation arteries and veins stream that with PNS is main component can improve the rat mesentery microcirculation disturbance that ischemia-reperfusion (I/R) causes, comprise suppress leukocyte sticking with the sticking of thin vein tube wall, thin vein wall peroxide generation and blood vessel around mast cell degranulation, PNS can also improve vascular endothelial function.The result of study proof gave the adhesion that PNS can suppress leukocyte and thin vein in our body in the past before LPS drops into, suppress mast cell degranulation in the body, results of in vitro studies has proved that PNS can suppress the expression of LPS inductive GA molecule CD18 and CD11b.
The main water soluble ingredient of the Radix Salviae Miltiorrhizae in the FUFANG DANSHEN DIWAN can be removed peroxide, suppress the expression of vascular endothelial cell adhesion molecule, and PNS can suppress the expression of LAM.The liver microcirculation disturbance that CP can improve ischemia-reperfusion has the improvement effect, suppresses the rat mesentery thrombosis that photochemical method causes.CP can also suppress the rat mesentery thrombosis that photochemical method causes in addition.But; whether the cardiac microcirculation disturbance of FUFANG DANSHEN DIWAN after to ischemia-reperfusion has the improvement effect; consideration convey to I-κ B-α degraded in the heart cell and P50 moves whether inhibitory action is arranged, to the Ultrastructural damage of myocardial cell whether protective effect etc. do not appear in the newspapers as yet.For this reason, the logical again ischemia-reperfusion injury model that causes after this experiment employing ligation left anterior descending coronary artery, dynamic observe thin arteriovenous diameter of rat heart coronary vasodilator and red cell velocity behind the ischemia-reperfusion with upright microscope that connects SIT video camera and high-speed camera and laser-Doppler blood flow instrument, the albuminous blood vessel of FITC labelling leaks outside and does well and the cardiac blood flow.With counting the myocardial infarction volume with the TTC staining; observe and count the myocardial cell of apoptosis with the TUNEL staining; Ultrastructural variation with transmission electron microscope observing cardiovascular and myocardial cell; detect the I-κ B-α of heart tissue with Western; P50, the expression of P65 is with the MPO in the uv-spectrophotometric instrument detection heart tissue; study FUFANG DANSHEN DIWAN to the protective effect of perfusion back cardiac microcirculation disturbance and myocardial damage again, and inquire into its mechanism.
Summary of the invention:
The invention provides the protective effect of FUFANG DANSHEN DIWAN to cardiac microcirculation disturbance and myocardial damage
FUFANG DANSHEN DIWAN of the present invention, Main Ingredients and Appearance are the compound Chinese medicinal preparation that Radix Salviae Miltiorrhizae, Radix Notoginseng and Borneolum Syntheticum are formed.
Red phenol total acid of the present invention, FUFANG DANSHEN DIWAN can have been bought from the market, also can meet medicinal standard and get final product according to prior art for preparing.
The present invention proves effect of the present invention by following experimental data
Experiment one
The rat heart microcirculation disturbance that the disposable pre-administration of FUFANG DANSHEN DIWAN causes ischemia-reperfusion and the preventive effect of myocardial damage
1. animal and reagent
Male SD rat, body weight 250 ± 10g buys the animal center in the Department Of Medicine, Peking University, the quality certification number: SCXK (capital) 2006-0001.The management of laboratory animal is pressed the management of laboratory animal guide of Ministry of Public Health issue and is carried out, and animal feeding is 24 ± 1 ℃ in temperature, and humidity is under 55 ± 5% the condition, free diet, drinking-water.
2. operation process
Rat is with 20% urethane (1.25g/kg) intramuscular anesthesia, and dorsal position is fixed, and the cervical region unhairing is cut skin, separates throat flesh, exposes trachea, the circulation of qi promoting cannula.Intubate end in addition is connected in the capable pressure breathing of toy respirator (respiratory quotient 1: 1,75 times/min of respiratory frequency, tidal volume 12mL/kg).(Ohio is USA) between the degree for YSI REF 401, Yellow Spring at 37-37.5 ℃ to keep the anus temperature with hot blanket.Chest unhairing, sterilization, along a breastbone left side, right border 2～4 ribs are opened breast and are exposed heart, cut off pericardium, pass following slightly (1～2) the mm place of pulmonary conus and left auricle boundary with being installed with sutural 3/8 looper of 3-0, the ligation stitching thread is also put a polyethylene tubule between silk thread and cardiac muscle group, the tension silk thread forms and causes ischemia, myocardium color to bleach to be the sign of ligation success.Loosen silk thread after 30 fens kinds and promptly take place to pour into again, it is again the successful sign of perfusion that ischemic area recovers redness, a sham operated rats not ligation of threading.
3. medicine and grouping
FUFANG DANSHEN DIWAN (Tianjin Tasly Pharmaceutical Co., Ltd provides, lot number: 20040502), preserve down by room temperature.The SD rat that experiment is selected for use, be divided into 5 groups (n=6) at random, sham operated rats (Sham), ischemia-reperfusion injury model (I/R) group, FUFANG DANSHEN DIWAN 0.1g/kg+ ischemia-reperfusion group (CP0.1+I/R), FUFANG DANSHEN DIWAN 0.4g/kg+ ischemia-reperfusion group (CP0.1+I/R), FUFANG DANSHEN DIWAN 0.8g/kg+ ischemia-reperfusion group (CP0.1+I/R).In experiment preceding 90 minutes, sham operated rats and ischemia-reperfusion group are irritated stomach respectively and are given normal saline 4ml/kg, each dosage group of FUFANG DANSHEN DIWAN is irritated stomach respectively and is given corresponding dosage FUFANG DANSHEN DIWAN (0.1g/kg, 0.4g/kg, 0.8g/kg) normal saline solution 4ml/kg.
4. heart coronaries blood vessel thin arteriovenous blood vessel footpath and red cell velocity
With being connected upright microscope (BX51WI, Olympus, (the ultimate APX PHOTRON of high-speed motion picture camera Japanese), FASTCAM US), is falling to penetrating under the light, 0 times of object lens of 1 usefulness, select the thin vein of diameter, by display screen (20PF5120, PHLIPS at 30-50 μ m, US) observe, and with CD videocorder (DVR-560H, PHLIPS, US) record.The picture rate of setting high-speed camera was 500 width of cloth/seconds, be recorded in ischemia before, ischemia 30 minutes, pour into again 30 minutes, pour into thin artery and vein caliber and erythrocyte flowing velocity in the 60 timesharing the same visual fields again.In the video recording of the speed playbacks of 25 width of cloth/seconds, measure thin artery and vein caliber and red cell velocity with Image-Pro Plus5.0 software.The thin artery and vein red cell velocity of rat heart is represented with mm/s, is basic value with the thin artery and vein red cell velocity before the ischemia-reperfusion, calculates the rate of change of respectively organizing the thin artery and vein red cell velocity of heart.The thin artery and vein caliber of rat heart is represented with mm, is basic value with the thin artery and vein caliber before the ischemia-reperfusion respectively, calculates the rate of change of the thin artery and vein caliber of heart.
5. the mensuration that spills of heart coronaries blood vessel thin vein plasma albumin
Other gets rat (every group of N=6), ischemia-reperfusion 60 minutes, the plasma albumin of FITC labelling is pressed the dosage of 50mg/kg, slowly injects through femoral vein.With just putting fluorescence microscope with the 455nm exciting light, mercury lamp is as transmitting illuminant (100W), when each group of record is poured into 60 minutes again in the thin vein blood vessel and the FITC fluorescent image of adjacent blood vessel outer room matter.Measure in the thin vein blood vessel and the FITC fluorescence intensity of adjacent blood vessel outer room matter with Image-Pro Plus 5.0 softwares, represent the value that rat heart thin vein albumin oozes out with the ratio of FITC fluorescence in the interior thin vein pipe outer room matter of the same visual field and the blood vessel.
6. the mensuration of cardiac blood flow
With the laser-Doppler blood perfusion imager that connects computer (PeriScan PIM3, PERIMED, Sweden) be recorded in ischemia before, ischemia 5 minutes, 10 minutes, 30 minutes and pour into 5 minutes, 10 minutes, 30 minutes, 60 timesharing heart surface blood flows again.With LDPIwin software image is measured and assessment data.With the blood flow before the ischemia-reperfusion is basic value, calculates the rate of change of cardiac blood flow.
7. the mensuration of myocardial infarction area
After pouring into 60 minutes again, take out rat heart (every group of N=6), begin to prolong from apex and be parallel to chamber direction at interval and be cut into 5 thick thin slices of 1mm and put into 0.375% TTC and under 37 ℃, hatched row TTC dyeing 15 minutes.Non-infarct is dyed redness, infarct is by white colouring, with Digital sight (DS-5M-U1, NIKON, Japan) take the myocardium picture of figure, (Media Cybemetrics Inc USA) calculates the percent value of the infarct size and the left ventricle area of every cardiac muscle, represents total infarct size with the meansigma methods of the percentage ratio of the myocardial infarction area of five hearts sections again with image analysis software Image-Pro plus5.0.
8. the TUNEL of myocardial cell dyeing
Irritating 90min again, get rat heart, perfusion fixation.Freezing section of cutting 6um, row TUNEL dyeing.With laser scanning co-focusing (Radiance2100 Bio-Rad UK), 63 times of object lens (Axiovert200Carl Zeiss Germany) are selected 5 visuals field in the heart ischemia reperfusion district, with image analysis software Image-Pro plus 5.0 (Media Cybemetrics Inc, USA) counting TUNEL positive cell number, TUNEL represents in positive cell number/visual field.
The ultrastructure of 9 heart blood capillaries and myocardial cell
Other gets part rat (every group of n=3), pour into 90 minutes again after, in the middle of ligature and the apex of the heart, get about 2mm
3Flesh tissue be used for electron microscopic observation.Tissue is with fixing before 3% glutaraldehyde, with fixing behind 1% the osmic acid.After sample dewaters with gradient acetone, with making super broad section after the epon 812 embedding.With acetic acid uranium and lead citrate dyeing transmission electron microscope (JEM-1230; JEOL Ltd., Tokyo Japan) observes heart blood capillary and myocardial cell ultrastructure.
10.MPO active detection
After pouring into 180 fens again, get the 100mg of cardiac muscular tissue in part Ischemia and Reperfusion in vivo in Rats zone, liquid nitrogen speed is cold, moves into-80 refrigerators.Add reactant mixtures extraction MPO at different levels after getting freezing heart homogenate, detect at the 460nm place, represent with the U/g heart tissue with the uv-spectrophotometric instrument.
11. the immunoblotting assay of I-κ B-α, P50, P65 in the rat heart muscle tissue
After pouring into 60 minutes again, get the 200mg of ishemic part cardiac muscular tissue, preserve in-80 refrigerators.With the protein extraction test kit whole protein and nucleoprotein (Applygen Technologies Inc.) are proposed.Sample thief adds isopyknic 2 * electrophoresis sample buffer mix after, behind the 10%SDS-PAGE electrophoresis (30mA 2hour RT.), electricity changes film to pvdf membrane (90mA spend the night ice bath).5% defatted milk powder sealing 1hour, 3 times * 5min of TBS-Tween rinsing, clip destination protein zone marker NF-κ B p65 (dilutes with 5% BSA-TBS-Tween 1:1000; SC-109-G, Santa Cruz), NF-κ B p50 (dilutes with 5% BSA-TBS-Tween 1:1000; SC-109-G, Santa Cruz), Tubulin (dilutes with 5% defatted milk powder 1:5000; 046k4770, sigma), I-κ B-α (dilutes with 5% BSA-TBS-Tween 1:1000; PC 142, Calbiochem Inc., and Germany) one is anti-, 4 ℃ of overnight incubation, Tbs-Tween washes 3 times * 5min, with two anti-hatching of peroxidase labelling, room temperature 1h, Tbs-Tween washes 3 times * 20min.The ECL 5min that develops the color, the magazine exposure.Colour developing band on the X-ray sheet carries out densitometric scan, with image analysis software (Image-Pro plus5.0, Media Cybemetrics Inc, USA) gray value of survey band.Gray value ratio with the I-κ B-α of Sham group and Tubulin is a basic value, calculates I-κ B-α and the ratio of Tubulin and the ratio of basic value of each group.With the P65 of Sham group or the gray value of P50 is basic value, calculates the P65 of each group or the gray value and basic value ratio of P50.
12. the mensuration of peripheral blood surfaces of granulocytes adhesion molecule expression
Each organizes rat after pouring into 60 minutes again, abdominal aortic blood, 3.8% sodium citrate anticoagulant (volume ratio of sodium citrate and blood is 1:9), mix rapidly in the 10ml centrifuge tube blood drawing back, get anticoagulated whole blood 50 μ l, the CD11b that adds 1 μ g FITC labelling, CD18 antibody (BD biosciences Pharmingen, USA), use the hemolysin broken red blood cell, after PBS washing 2 times, with flow cytometer (FACS Calibur, BD, USA) according to forward angle/lateral angle sorting granulocyte, count 5000 granulocytes, measure the anti-CD11b of FITC labelling and the positive cell number and the average fluorescent strength of anti-CD18 antibody.
13. statistical method
Statistical analysis adopts SPSS 11.5 mathematical statistics software kits, and each is organized data and adds up with One-Way ANOVA, carries out the multiple comparisons analysis with S-N-K (variance is neat) or Tamhane ' s T2 (heterogeneity of variance) when each group relatively has statistical significance.Data represent with x ± SE, represent that difference has statistical significance when P＜0.05.
The result
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the influence of the thin arteriovenous caliber of rat heart coronary vasodilator
Figure 1A represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator arteriole caliber.Sham group rat is in this viewing duration, and rat heart coronary vasodilator arteriole caliber does not change significantly.I/R group rat is early stage at ischemia-reperfusion, and heart coronaries blood vessel arteriole caliber has the property a crossed contraction, and the pre-administration of CP0.1, CP0.4 and CP0.8 suppresses to pour into the property crossed a contraction contraction of early stage rat heart coronary vasodilator arteriole significantly again.
Figure 1B represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator thin vein caliber.I/R group rat heart thin vein caliber does not change in this viewing duration significantly, and each pre-administration group rat heart thin vein caliber of CP is not significant yet to be changed.
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the influence of the thin arteriovenous red cell velocity of rat heart coronary vasodilator
Fig. 2 A represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator arteriole red cell velocity.The crown arteriole red cell velocity of Sham group rat does not change in this viewing duration significantly.I/R group rat coronary vasodilator arteriole red cell velocity reduces when perfusion begins again significantly, pours into after 30 minutes again and recovers.The pre-administration of CP0.1 and CP0.4 does not have remarkable influence to the crown arteriole red cell velocity of rat.The CP0.8+I/R group is from pouring into 30 again, and rat coronary vasodilator arteriole red cell velocity is higher than the I/R group significantly.
Fig. 2 B represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator thin vein red cell velocity.The crown thin vein red cell velocity of Sham group rat does not change in this viewing duration significantly.The just reduction significantly when perfusion begins again of I/R group rat coronary vasodilator thin vein red cell velocity, and last till when this observation finishes.CP0.1+I/R group and CP0.1+I/R group group rat suppress to pour into the reduction of the rat coronary vasodilator thin vein red cell velocity that causes significantly again, and CP0.8+I/R group rat, from pouring into again 30 fens, suppressed the reduction of the red cell velocity that ischemia-reperfusion causes significantly, and held to continue and pour into 60 fens again.
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the influence of oozing out of rat heart coronary vasodilator thin vein plasma albumin
Fig. 3 A represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the albuminous state that spills of CP0.8+I/R group rat coronary vasodilator thin vein when pouring into 60 minutes again.The blood vessel that Sham group rat only can observe the small amount of FITC tagged albumin spills (A-1) outward.The blood vessel that I/R group rat can obviously observe the FITC tagged albumin spills (A-2) outward.The blood vessel of the pre-administration group rat heart coronary vasodilator thin vein FITC tagged albumin of CP0.1 and CP0.4 leaks outside out, and (A-3, A-4), the blood vessel of the FITC tagged albumin of CP0.8+I/R group rat spills obvious minimizing (A-5) outward in not minimizing.
Fig. 3 B is illustrated in the comparison of Sham group when irritating 60 minutes again, I/R group, CP0.1+I/R group, CP0.4+I/R group, the inside and outside FITC fluorescence intensity ratio of CP0.8+I/R group rat heart coronary vasodilator thin vein.Sham group rat heart thin vein blood vessel outer with blood vessel in the ratio of the optical density that spills of FITC tagged albumin be 20.13 ± 2.52.The ratio of I/R group rat is 41.44 ± 2.46, is significantly higher than sham operated rats.CP0.1+I/R group and CP0.4+I/R group rat heart thin vein blood vessel the ratio outer and optical density that the interior FITC tagged albumin of blood vessel spills is respectively 37.42 ± 3.61,33.46 ± 3.32 and does not suppress albuminous spilling significantly.CP0.8+I/R group rat heart thin vein blood vessel the ratio outer and luminosity that the interior FITC tagged albumin of blood vessel spills is 31.23 ± 3.75, significantly is lower than ischemia-reperfusion group.
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the influence of rat heart blood flow
What Fig. 4 represented is that Sham group, I/R group, CP0.1+I/R group, the CP0.4+I/R that the laser-Doppler survey obtains organizes, the image of CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have obvious variation (A) in this process.I/R group rat blood flow when pouring into 30min and 60min more obviously reduce (B3, B4).CP0.1+I/R group, CP0.4+I/R group rat heart blood flow when pouring into 30min again, compare to some extent and recover with I/R with 60min (C, D).CP0.8+I/R group rat cardiac blood flow when pouring into 30min and 60min more obviously recovers (E).
The continuous variation of the expression Sham group of Fig. 5, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have significant change in this observation process.I/R group rat heart blood flow is in ischemia and reduction significantly between flush phase again, and the pre-administration of CP0.1 and CP0.4 does not suppress the reduction of the rat heart blood flow behind the ischemia-reperfusion significantly.CP0.8+I/R group from pour into again 5 assign to pour into again 30 minutes during in, suppressed the reduction of the rat heart blood flow that perfusion again causes significantly.
FUFANG DANSHEN DIWAN to rat myocardial ischemia and reperfusion after the influence of myocardial infarction area
Fig. 6 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart TTC colored graph when CP0.8+I/R group rat is poured into 60 minutes again.Sham group rat heart is not seen infarcted region (A), the visible bigger painted infarcted region (B) that is white in color of I/R group rat heart.The myocardial infarction zone of CP0.1+I/R group, CP0.4+I/R group reduces slightly that (C, D), (E) reduced in CP0.8+I/R group myocardial infarction zone significantly.
Fig. 7 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, when CP0.8+I/R group rat is poured into 60 minutes again the heart myocardial infarction area and.I/R organizes rat heart muscle infarcted region area and is 31.43 ± 4.07mm
3CP0.1+I/R group and CP0.4+I/R group rat heart muscle infarct size be respectively 26.63 ± 4.85,24.58 ± 5.3mm
3, do not reduce myocardial infarction area significantly.CP0.8+I/R organizes the rat heart muscle infarct size and is 22.55 ± 1.9mm
3, reduced the myocardial infarction area that ischemia-reperfusion causes significantly.
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the influence of apoptosis of cardiac muscle
Fig. 8 A is that Sham group, I/R organize, CP0.1+I/R organizes, CP0.4+I/R organizes, CP0.8+I/R group rat is poured into 90 o'clock painted images of heart TUNEL again.Sham organizes the male myocardial cell of the accidental TUNEL of rat (A-1), I/R group rat heart can be observed the male myocardial cell of a large amount of TUNEL (A-2), the all minimizings significantly of the TUNEL positive cardiomyocytes of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group (A-3, A-4, A-5).
Fig. 8 B represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart TUNEL positive cardiomyocytes result.Sham group rat TUNEL positive cardiomyocytes only is 0.36 ± 0.11/visual field, and I/R group rat TUNEL positive cardiomyocytes is 14.7 ± 1.17/visual field, significantly organizes more than Sham.CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat TUNEL positive cardiomyocytes quantity are respectively 9.9 ± 0.71,7.16 ± 1.431 and 5.56 ± 1 (individual/visual field), all are markedly inferior to the I/R group.
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after the Ultrastructural influence of rat heart
Fig. 9 A represents the transmission electron microscope observing result of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart blood capillary.Sham group rat heart capillary endothelium is complete, can be observed a spot of vesicle, does not observe the edema (A-1) of blood vessel periphery.The capillary endothelial cell swelling of I/R group rat heart can be observed big pinosome and blood vessel periphery edema (A-2) in the vascular endothelial cell.The capillary endothelial cell swelling of CP0.1+I/R group, CP0.4+I/R group rat heart alleviates, do not observe big pinosome in the endotheliocyte and blood vessel periphery decrease in edema (A-3, A-4).The capillary endothelial cell of CP0.8+I/R group rat heart is intact, does not see endotheliocytic swelling and blood vessel periphery edema (A-5).
Fig. 9 B represents the projection electron microscopic observation result of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat myocardial cell.Sham group rat heart myocardial cell fiber alignment is neat, evenly, closely arrangement of mitochondrion, regular shape, peplos are complete, the dense collection of ridge, rule (B-1).I/R rat heart myocardial cell edema, myofilament dissolving, fracture, mitochondrial swelling, many places vacuolation (B-2).CP concentration has been improved the ultrastructure of heart myocardial cell interdependently, and fiber alignment is more neat, and mitochondrial swelling alleviates, and peplos is complete, and the form rule (B-3, B-4, B-5).
FUFANG DANSHEN DIWAN to myocardial ischemia-reperfusion after rat heart muscle organize the influence of MPO content
Figure 10 shows Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart left ventricular ischemia content of perfusion area tract tissue MPO again.The value of Sham group MPO is 0.18 ± 0.016U/g cardiac muscular tissue, the MPO value of I/R group rises to 0.63 ± 0.034U/g cardiac muscular tissue, the MPO value of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group is respectively 0.55 ± 0.027,0.54 ± 0.026 and 0.5 ± 0.022 (U/g cardiac muscular tissue), the inhibition of concentration interdependence the rat heart left ventricular ischemia pour into the content of MPO in the regional cardiac muscular tissue again.
FUFANG DANSHEN DIWAN to ischemia-reperfusion after the influence of I-κ B-α in the rat heart tissue
When Figure 11 A represents to pour into 60 minutes again, the immunoblotting assay image of I-κ B-a in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with Sham group, I/R group rat heart organizes I-κ B-a band gray scale obviously to reduce, and the pre-administration of CP0.8g/kg has suppressed the minimizing that rat heart that ischemia-reperfusion causes is organized I-κ B-a band gray scale.
When Figure 11 B has represented to pour into 60 minutes again, the result of the Weatern blot quantitative analysis that whole protein I-κ B-a expresses in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with the Sham group, I/R group rat heart is organized I-κ B-a/Tubulin gray scale than obviously reducing, and CP0.1+I/R group, CP0.4+I/R group I-κ B-a/Tubulin gray scale are not suppressed significantly than reducing.The pre-administration of CP0.8g/kg has suppressed the minimizing that rat heart that ischemia-reperfusion causes is organized I-κ B-a/Tubulin gray scale ratio.
FUFANG DANSHEN DIWAN to ischemia-reperfusion after rat heart organize the influence of P50, P65
When Figure 12 A represents to pour into 60 minutes again, the immunoblotting assay image of NF-κ B subunit P50 and P65 in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with the Sham group, I/R group nucleoprotein P50 band gray scale when pouring into 60 minutes more obviously increases.The nucleoprotein P50 band gray scale of CP0.4+I/R group, CP0.8+I/R group obviously reduces.Yet I/R organizes when pouring into 60 minutes again, does not detect the band of nucleoprotein P65.Each dosage group of CP does not detect the band of nucleoprotein P65 yet when pouring into 60 minutes again.
The Western Blotting analysis result of the Sham group that Figure 12 B shows, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat rat heart nucleus albumen P50 and P65 protein content.Compare with Sham group rat, I/R organizes after pouring into 60 fens again, and the gray scale of rat heart nucleus egg albumen P50 significantly increases, and the proteic consideration convey of prompting P50 moves increase.The pre-administration of CP0.1g/kg does not have significantly that the gray scale of rat heart nucleus albumen P50 significantly increases, and the pre-administration of CP0.4g/kg and CP0.8g/kg has suppressed significantly behind the ischemia-reperfusion that the proteic consideration convey of P50 moves in the rat heart tissue.But, compare with Sham group rat, not significant variation of I/R group rat heart nucleus albumen P65, the rat heart nucleus albumen P65 of each pre-administration group of CP is not significant yet to be changed.
11. FUFANG DANSHEN DIWAN is brought out the influence of adhesion molecule to myocardial ischemia reperfusion injury
Figure 13 A represents respectively to organize the variation of leukocyte adhesion factor CD11b, and sham operated rats leukocyte adhesion molecule CD11b average fluorescent strength is 31.11 ± 1.23.The leukocyte adhesion molecule CD11b average fluorescent strength of ischemia-reperfusion group rises to 41.86 ± 2.87, with the sham operated rats comparison and have statistical significance.The fluorescence intensity level that FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into back leukocyte adhesion factor CD11b in advance is respectively 40.66 ± 2.87,42.7 ± 3.9 and 35.07 ± 1.35.From as a result we as can be seen FUFANG DANSHEN DIWAN drop in advance the back CD11b expression there is no obvious influence.
Figure 13 B represents respectively to organize the variation of leukocyte adhesion factor CD18, and sham operated rats leukocyte adhesion molecule CD18 average fluorescent strength is 49.1 ± 3.59.The leukocyte adhesion molecule CD18 average fluorescent strength of ischemia-reperfusion group rises to 68.93 ± 4.21, with the sham operated rats comparison and have statistical significance.The fluorescence intensity level that FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into back leukocyte adhesion factor CD18 in advance is respectively 74.38 ± 4.83,73.59 ± 4.93 and 57.99 ± 3.8.From as a result we as can be seen FUFANG DANSHEN DIWAN 0.8g/kg can obviously suppress the expression of the leukocyte surface adhesion molecule CD18 that ischemia-reperfusion causes.
Brief summary
1. the disposable pre-administration of the FUFANG DANSHEN DIWAN 0.8g/kg rat heart microcirculation disturbance that can prevent ischemia-reperfusion to cause, comprise the contraction that suppresses rat coronary vasodilator arteriole, blood flow rate reduces, and albumin spills increase, the reduction of blood flow reduction and myocardial flow.
2. the rat heart muscle infarction gross area that causes of FUFANG DANSHEN DIWAN 0.8g/kg disposable pre-administration can minimizing ischemia-reperfusion.
3. the inhibition myocardial cell that causes of FUFANG DANSHEN DIWAN 0.4g/kg, 0.8g/kg disposable pre-administration can minimizing ischemia-reperfusion is transferred and is died.
4. heart tissue MPO content increased after FUFANG DANSHEN DIWAN .0.1g/kg, 0.4g/kg, the disposable pre-administration of 0.8g/kg all can suppress ischemia-reperfusion.
5. the disposable pre-administration of FUFANG DANSHEN DIWAN 0.8g/kg can suppress the expression of peripheral blood GA molecule CD18 behind the ischemia-reperfusion.
6. interior I-κ B-a degraded and the P50 consideration convey of the disposable pre-administration inhibition ischemia-reperfusion rear myocardium tissue's cytoplasm of FUFANG DANSHEN DIWAN 0.8g/kg moves increase.
Conclusion
The disposable pre-input of this result of study prompting FUFANG DANSHEN DIWAN 0.8g/kg can be improved ischemia-reperfusion rat microcirculation disturbance and myocardial damage.This effect may with the expression of its inhibition GA molecule CD18, suppress the activation of MPO, the consideration convey of I-κ B-a degraded and P50 moves, and suppresses the myocardial cell accent and dies relevant.The rat heart microcirculation disturbance that the repeatedly pre-medicine of FUFANG DANSHEN DIWAN causes ischemia-reperfusion and the protective action of myocardial damage.
Experiment two: the rat heart microcirculation disturbance that the repeatedly pre-medicine of FUFANG DANSHEN DIWAN causes ischemia-reperfusion and the protective action of myocardial damage
Experimental technique is identical with experiment one with material in the experiment two
1. animal and reagent
Male SD rat, body weight 250 ± 10g buys the animal center in the Department Of Medicine, Peking University, the quality certification number: SCXK (capital) 2006-0001.The management of laboratory animal is pressed the management of laboratory animal guide of Ministry of Public Health issue and is carried out, and animal feeding is 24 ± 1 ℃ in temperature, and humidity is under 55 ± 5% the condition, free diet, drinking-water.
2. operation process
Rat is with 20% urethane (1.25g/kg) intramuscular anesthesia, and dorsal position is fixed, and the cervical region unhairing is cut skin, separates throat flesh, exposes trachea, the circulation of qi promoting cannula.Intubate end in addition is connected in the capable pressure breathing of toy respirator (respiratory quotient 1: 1,75 times/min of respiratory frequency, tidal volume 12mL/kg).(Ohio is USA) between the degree for YSI REF 401, Yellow Spring at 37-37.5 ℃ to keep the anus temperature with hot blanket.Chest unhairing, sterilization, along a breastbone left side, right border 2～4 ribs are opened breast and are exposed heart, cut off pericardium, pass following slightly (1～2) the mm place of pulmonary conus and left auricle boundary with being installed with sutural 3/8 looper of 3-0, the ligation stitching thread is also put a polyethylene tubule between silk thread and cardiac muscle group, the tension silk thread forms and causes ischemia, myocardium color to bleach to be the sign of ligation success.Loosen silk thread after 30 fens kinds and promptly take place to pour into again, it is again the successful sign of perfusion that ischemic area recovers redness, a sham operated rats not ligation of threading.
3. medicine and grouping
FUFANG DANSHEN DIWAN (Tianjin Tasly Pharmaceutical Co., Ltd provides, lot number: 20040502), preserve down by room temperature.The SD rat that experiment is selected for use is divided into 5 groups (n=6) at random.
Sham operated rats (Sham): irritate stomach every day and give normal saline 4ml/kg, continuous 6 days.
Ischemia-reperfusion injury model (I/R) group: irritate stomach every day and give normal saline 4ml/kg, continuous 6 days.
Continuous 6 days administration+ischemia-reperfusion group (CP0.1+I/R) of FUFANG DANSHEN DIWAN 0.1g/kg: irritate stomach every day and give FUFANG DANSHEN DIWAN 0.1g/kg, continuous 6 days.
Continuous 6 days administration+ischemia-reperfusion group (CP0.4+I/R) of FUFANG DANSHEN DIWAN 0.4g/kg: irritate stomach every day and give FUFANG DANSHEN DIWAN 0.4g/kg, continuous 6 days.
Continuous 6 days administration+ischemia-reperfusion group (CP0.8+I/R) of FUFANG DANSHEN DIWAN 0.8g/kg: irritate stomach every day and give FUFANG DANSHEN DIWAN 0.8g/kg, continuous 6 days.
4. the mensuration of cardiac blood flow
With the laser-Doppler blood perfusion imager that connects computer (PeriScan PIM3, PERIMED, Sweden) be recorded in ischemia before, ischemia 5 minutes, 10 minutes, 30 minutes and pour into 5 minutes, 10 minutes, 30 minutes, 60 timesharing heart surface blood flows again.With LDPIwin software image is measured and assessment data.With the blood flow before the ischemia-reperfusion is basic value, calculates the rate of change of cardiac blood flow.
5. the mensuration of myocardial infarction area
After pouring into 60 minutes again, take out rat heart (every group of N=6), begin to prolong from apex and be parallel to chamber direction at interval and be cut into 5 thick thin slices of 1mm and put into 0.375% TTC and under 37 ℃, hatched row TTC dyeing 15 minutes.Non-infarct is dyed redness, infarct is by white colouring, with Digital sight (DS-5M-U1, NIKON, Japan) take the myocardium picture of figure, (Media Cybemetrics Inc USA) calculates the percent value of the infarct size and the left ventricle area of every cardiac muscle, represents total infarct size with the meansigma methods of the percentage ratio of the myocardial infarction area of five hearts sections again with image analysis software Image-Pro plus5.0.
6. the TUNEL of myocardial cell dyeing
Irritating 90min again, get rat heart, perfusion fixation.Freezing section of cutting 6um, row TUNEL dyeing.With laser scanning co-focusing (Radiance2100 Bio-Rad UK), 63 times of object lens (Axiovert200Carl Zeiss Germany) are selected 5 visuals field in the heart ischemia reperfusion district, with image analysis software Image-Pro plus 5.0 (Media Cybemetrics Inc, USA) counting TUNEL positive cell number, TUNEL represents in positive cell number/visual field.
7. the expression of peripheral blood GA molecule CD11b/CD18
Each organizes rat after pouring into 60 minutes again, abdominal aortic blood, 3.8% sodium citrate anticoagulant (volume ratio of sodium citrate and blood is 1:9), mix rapidly in the 10ml centrifuge tube blood drawing back, get anticoagulated whole blood 50 μ l, the CD11b that adds 1 μ g FITC labelling, CD18 antibody (BD biosciences Pharmingen, USA), use the hemolysin broken red blood cell, after PBS washing 2 times, with flow cytometer (FACS Calibur, BD, USA) according to forward angle/lateral angle sorting granulocyte, count 5000 granulocytes, measure the anti-CD11b of FITC labelling and the positive cell number and the average fluorescent strength of anti-CD18 antibody.
8. statistical method
Statistical analysis adopts SPSS11.5 mathematical statistics software kit, and each is organized data and adds up with One-Way ANOVA, carries out the multiple comparisons analysis with S-N-K (variance is neat) or Tamhane ' s T2 (heterogeneity of variance) when each group relatively has statistical significance.Data represent with x ± SE, represent that difference has statistical significance when P＜0.05.
The result
The FUFANG DANSHEN DIWAN successive administration to myocardial ischemia-reperfusion after the influence of rat heart blood flow
What Figure 14 represented is that Sham group, I/R group, CP0.1+I/R group, the CP0.4+I/R that laser-Doppler records organizes, the image of CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have obvious variation in this process.I/R group rat blood flow after pouring into again obviously reduces.CP0.1+I/R group rat heart blood flow is compared to some extent with I/R when pouring into 30min with 60min again and is recovered.CP0.4+I/R group, CP0.8+I/R group rat cardiac blood flow after pouring into again obviously recover.
The continuous variation of the expression Sham group of Figure 15, I/R group, CP0.1+I/R, CP0.4+I/R group, CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have significant change in this observation process.I/R group rat heart blood flow is in ischemia and reduction significantly between flush phase again, and the CP0.1g/kg successive administration does not suppress the reduction of the rat heart blood flow behind the ischemia-reperfusion significantly.The reduction that can significantly suppress to pour into again the blood flow that causes in 60 minutes was being poured into 5 fens and poured into to the CP0.4+I/R group again.CP0.8+I/R group from pour into again 5 assign to pour into again 60 minutes during in, all can significantly suppress to pour into again the reduction of the blood flow that causes.The FUFANG DANSHEN DIWAN successive administration to rat myocardial ischemia and reperfusion after the influence of myocardial infarction area
Figure 16 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart TTC colored graph when CP0.8+I/R group rat is poured into 60 minutes again.Sham group rat heart is not seen infarcted region, the visible bigger painted infarcted region that is white in color of I/R group rat heart.Reduce significantly in CP0.1+I/R group, CP0.4+I/R group and CP0.8+I/R group myocardial infarction zone.
Figure 17 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart myocardial infarction area statistic analysis result when CP0.8+I/R group rat is poured into 60 minutes again.I/R group rat heart muscle infarcted region area is 15.55 ± 2.14%.CP0.1+I/R group, CP0.4+I/R group and CP0.8+I/R group rat heart muscle infarct size be respectively 8.32 ± 1.42%, 7.65 ± 1.66%, 7.56 ± 2.57%, reduced the area of the myocardial infarction that ischemia-reperfusion causes significantly.
The FUFANG DANSHEN DIWAN successive administration to myocardial ischemia-reperfusion after the influence of apoptosis of cardiac muscle
Figure 18 pours into 90 o'clock painted images of heart TUNEL again for Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat.The male myocardial cell of the Sham group accidental TUNEL of rat, I/R group rat heart can be observed the male myocardial cell of a large amount of TUNEL, and the TUNEL positive cardiomyocytes of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group all reduces significantly.
Figure 19 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart TUNEL positive cardiomyocytes result.Sham group rat TUNEL positive cardiomyocytes only is 0.36 ± 0.11/visual field, and I/R group rat TUNEL positive cardiomyocytes is 14.7 ± 1.17/visual field, significantly organizes more than Sham.CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat TUNEL positive cardiomyocytes quantity are respectively 7.1 ± 0.54,7.23 ± 0.66 and 6.4 ± 0.44 (individual/visual field), all are markedly inferior to the I/R group.
The FUFANG DANSHEN DIWAN successive administration to myocardial ischemia-reperfusion after the influence of peripheral blood GA developed by molecule
Figure 20 A represents respectively to organize the variation of peripheral blood GA molecule CD11b.Sham operated rats peripheral blood GA molecule CD11b average fluorescent strength is 31.11 ± 1.23.Ischemia-reperfusion group peripheral blood GA molecule CD11bCD11b average fluorescent strength rises to 41.86 ± 2.87, is higher than sham operated rats significantly.FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into the back does not in advance have significant inhibitory effect to the increase of the expression of GA factor CD11b.
Figure 20 B represents respectively to organize the variation of leukocyte adhesion factor CD18, and sham operated rats leukocyte adhesion molecule CD18 average fluorescent strength is 49.1 ± 3.59.The leukocyte adhesion molecule CD18 average fluorescent strength of ischemia-reperfusion group rises to 68.93 ± 4.21, is higher than sham operated rats significantly.FUFANG DANSHEN DIWAN 0.1g/kg, be respectively 54.15 ± 2.09 0.4g/kg drop into the fluorescence intensity level of back leukocyte adhesion factor CD18 in advance with 0.8g/kg, 57.42 ± 2.16 and 56.73 ± 2.79, suppressed the expression of the leukocyte surface adhesion molecule CD18 that ischemia-reperfusion causes significantly.
Brief summary
The rat heart blood flow reduces behind the ischemia-reperfusion, and the myocardial cell number of heart infarct size, apoptosis increases significantly, and peripheral blood cells adhesion molecule CD11b/CD18 expresses and increases significantly.0.1g/kg, the repeatedly pre-administration of FUFANG DANSHEN DIWAN of 0.4g/kg and 0.8g/kg consumption can dosage suppresses the reduction of the rat heart blood flow behind the ischemia-reperfusion interdependently, the myocardial cell number of heart infarct size and apoptosis suppresses the expression of peripheral blood granulocyte adhesion molecule CD18 significantly.
Conclusion
The muptiple-use pre-administration of FUFANG DANSHEN DIWAN can dosage prevents the reduction of the rat heart blood flow that ischemia-reperfusion causes interdependently, suppresses the damage of myocardial cell.
Description of drawings
Figure 1A represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator arteriole caliber.Sham group rat is in this viewing duration, and rat heart coronary vasodilator arteriole caliber does not change significantly.I/R group rat is early stage at ischemia-reperfusion, and heart coronaries blood vessel arteriole caliber has the property a crossed contraction, and the pre-administration of CP0.1, CP0.4 and CP0.8 suppresses to pour into the property crossed a contraction contraction of early stage rat heart coronary vasodilator arteriole significantly again.
Figure 1B represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator thin vein caliber.I/R group rat heart thin vein caliber does not change in this viewing duration significantly, and each pre-administration group rat heart thin vein caliber of CP is not significant yet to be changed.
Fig. 2 A represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator arteriole red cell velocity.The crown arteriole red cell velocity of Sham group rat does not change in this viewing duration significantly.I/R group rat coronary vasodilator arteriole red cell velocity reduces when perfusion begins again significantly, pours into after 30 minutes again and recovers.The pre-administration of CP0.1 and CP0.4 does not have remarkable influence to the crown arteriole red cell velocity of rat.The CP0.8+I/R group is from pouring into 30 again, and rat coronary vasodilator arteriole red cell velocity is higher than the I/R group significantly.
Fig. 2 B represents the continuous variation of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat coronary vasodilator thin vein red cell velocity.The crown thin vein red cell velocity of Sham group rat does not change in this viewing duration significantly.The just reduction significantly when perfusion begins again of I/R group rat coronary vasodilator thin vein red cell velocity, and last till when this observation finishes.CP0.1+I/R group and CP0.1+I/R group group rat suppress to pour into the reduction of the rat coronary vasodilator thin vein red cell velocity that causes significantly again, and CP0.8+I/R group rat, from pouring into again 30 fens, suppressed the reduction of the red cell velocity that ischemia-reperfusion causes significantly, and held to continue and pour into 60 fens again.
Fig. 3 A represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the albuminous state that spills of CP0.8+I/R group rat coronary vasodilator thin vein when pouring into 60 minutes again.Sham group rat only can
Spill (A-1) outward with the blood vessel of observing the small amount of FITC tagged albumin.The blood vessel that I/R group rat can obviously observe the FITC tagged albumin spills (A-2) outward.The blood vessel of the pre-administration group rat heart coronary vasodilator thin vein FITC tagged albumin of CP0.1 and CP0.4 leaks outside out, and (A-3, A-4), the blood vessel of the FITC tagged albumin of CP0.8+I/R group rat spills obvious minimizing (A-5) outward in not minimizing.
Fig. 3 B is illustrated in the comparison of Sham group when irritating 60 minutes again, I/R group, CP0.1+I/R group, CP0.4+I/R group, the inside and outside FITC fluorescence intensity ratio of CP0.8+I/R group rat heart coronary vasodilator thin vein.Sham group rat heart thin vein blood vessel outer with blood vessel in the ratio of the optical density that spills of FITC tagged albumin be 20.13
±2.52。The ratio of I/R group rat is 41.44 ± 2.46, is significantly higher than sham operated rats.CP0.1+I/R group and CP0.4+I/R group rat heart thin vein blood vessel the ratio outer and optical density that the interior FITC tagged albumin of blood vessel spills is respectively 37.42 ± 3.61,33.46 ± 3.32 and does not suppress albuminous spilling significantly.CP0.8+I/R group rat heart thin vein blood vessel the ratio outer and luminosity that the interior FITC tagged albumin of blood vessel spills is 31.23 ± 3.75, significantly is lower than ischemia-reperfusion group.
What Fig. 4 represented is that Sham group, I/R group, CP0.1+I/R group, the CP0.4+I/R that the laser-Doppler survey obtains organizes, the image of CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have obvious variation (A) in this process.I/R group rat blood flow when pouring into 30min and 60min more obviously reduce (B3, B4).CP0.1+I/R group, CP0.4+I/R group rat heart blood flow when pouring into 30min again, compare to some extent and recover with I/R with 60min (C, D).CP0.8+I/R group rat cardiac blood flow when pouring into 30min and 60min more obviously recovers (E).
The continuous variation of the expression Sham group of Fig. 5, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have significant change in this observation process.I/R group rat heart blood flow is in ischemia and reduction significantly between flush phase again, and the pre-administration of CP0.1 and CP0.4 does not suppress the reduction of the rat heart blood flow behind the ischemia-reperfusion significantly.CP0.8+I/R group from pour into again 5 assign to pour into again 30 minutes during in, suppressed the reduction of the rat heart blood flow that perfusion again causes significantly.
Fig. 6 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart TTC colored graph when CP0.8+I/R group rat is poured into 60 minutes again.Sham group rat heart is not seen infarcted region (A), the visible bigger painted infarcted region (B) that is white in color of I/R group rat heart.The heart of CP0.1+I/R group, CP0.4+I/R group
The flesh infarcted region reduces slightly that (C, D), (E) reduced in CP0.8+I/R group myocardial infarction zone significantly.
Fig. 7 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, when CP0.8+I/R group rat is poured into 60 minutes again the heart myocardial infarction area and.I/R organizes rat heart muscle infarcted region area and is 31.43 ± 4.07mm
3CP0.1+I/R group and CP0.4+I/R group rat heart muscle infarct size be respectively 26.63 ± 4.85,24.58 ± 5.3mm
3, do not reduce myocardial infarction area significantly.CP0.8+I/R organizes the rat heart muscle infarct size and is 22.55 ± 1.9mm
3, reduced the myocardial infarction area that ischemia-reperfusion causes significantly.
Fig. 8 A is that Sham group, I/R organize, CP0.1+I/R organizes, CP0.4+I/R organizes, CP0.8+I/R group rat is poured into 90 o'clock painted images of heart TUNEL again.Sham organizes the male myocardial cell of the accidental TUNEL of rat (A-1), I/R group rat heart can be observed the male myocardial cell of a large amount of TUNEL (A-2), the all minimizings significantly of the TUNEL positive cardiomyocytes of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group (A-3, A-4, A-5).
Fig. 8 B represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart TUNEL positive cardiomyocytes result.Sham group rat TUNEL positive cardiomyocytes only is 0.36 ± 0.11/visual field, and I/R group rat TUNEL positive cardiomyocytes is 14.7 ± 1.17/visual field, significantly organizes more than Sham.CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat TUNEL positive cardiomyocytes quantity are respectively 9.9 ± 0.71,7.16 ± 1.431 and 5.56 ± 1 (individual/visual field), all are markedly inferior to the I/R group.
Fig. 9 A represents the transmission electron microscope observing result of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart blood capillary.Sham group rat heart capillary endothelium is complete, can be observed a spot of vesicle, does not observe the edema (A-1) of blood vessel periphery.The capillary endothelial cell swelling of I/R group rat heart can be observed big pinosome and blood vessel periphery edema (A-2) in the vascular endothelial cell.The capillary endothelial cell swelling of CP0.1+I/R group, CP0.4+I/R group rat heart alleviates, do not observe big pinosome in the endotheliocyte and blood vessel periphery decrease in edema (A-3, A-4).The capillary endothelial cell of CP0.8+I/R group rat heart is intact, does not see endotheliocytic swelling and blood vessel periphery edema (A-5).
Fig. 9 B represents the projection electron microscopic observation result of Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat myocardial cell.Sham group rat heart myocardial cell fiber alignment is neat, evenly, closely arrangement of mitochondrion, regular shape, peplos are complete, the dense collection of ridge, rule (B-1).I/R rat heart myocardial cell edema, myofilament dissolving, fracture, mitochondrial swelling, many places vacuolation (B-2).CP concentration has been improved the ultrastructure of heart myocardial cell interdependently, and fiber alignment is more neat, and mitochondrial swelling alleviates, and peplos is complete, and the form rule (B-3, B-4, B-5).
Figure 10 shows Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart left ventricular ischemia content of perfusion area tract tissue MPO again.The value of Sham group MPO is 0.18 ± 0.016U/g cardiac muscular tissue, the MPO value of I/R group rises to 0.63 ± 0.034U/g cardiac muscular tissue, the MPO value of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group is respectively 0.55 ± 0.027,0.54 ± 0.026 and 0.5 ± 0.022 (U/g cardiac muscular tissue), the inhibition of concentration interdependence the rat heart left ventricular ischemia pour into the content of MPO in the regional cardiac muscular tissue again.
When Figure 11 A represents to pour into 60 minutes again, the immunoblotting assay image of I-κ B-a in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with Sham group, I/R group rat heart organizes I-κ B-a band gray scale obviously to reduce, and the pre-administration of CP0.8g/kg has suppressed the minimizing that rat heart that ischemia-reperfusion causes is organized I-κ B-a band gray scale.
When Figure 11 B has represented to pour into 60 minutes again, the result of the Weatern blot quantitative analysis that whole protein I-κ B-a expresses in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with the Sham group, I/R group rat heart is organized I-κ B-a/Tubulin gray scale than obviously reducing, and CP0.1+I/R group, CP0.4+I/R group I-κ B-a/Tubulin gray scale are not suppressed significantly than reducing.The pre-administration of CP0.8g/kg has suppressed the minimizing that rat heart that ischemia-reperfusion causes is organized I-κ B-a/Tubulin gray scale ratio.
When Figure 12 A represents to pour into 60 minutes again, the immunoblotting assay image of NF-κ B subunit P50 and P65 in Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, the CP0.8+I/R group rat heart tissue.Compare with the Sham group, I/R group nucleoprotein P50 band gray scale when pouring into 60 minutes more obviously increases.The nucleoprotein P50 band gray scale of CP0.4+I/R group, CP0.8+I/R group obviously reduces.Yet I/R organizes when pouring into 60 minutes again, does not detect the band of nucleoprotein P65.Each dosage group of CP does not detect the band of nucleoprotein P65 yet when pouring into 60 minutes again.
The Western Blotting analysis result of the Sham group that Figure 12 B shows, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat rat heart nucleus albumen P50 and P65 protein content.Compare with Sham group rat, I/R organizes after pouring into 60 fens again, and the gray scale of rat heart nucleus egg albumen P50 significantly increases, and the proteic consideration convey of prompting P50 moves increase.The pre-administration of CP0.1g/kg does not have significantly that the gray scale of rat heart nucleus albumen P50 significantly increases, and the pre-administration of CP0.4g/kg and CP0.8g/kg has suppressed significantly behind the ischemia-reperfusion that the proteic consideration convey of P50 moves in the rat heart tissue.But, compare with Sham group rat, not significant variation of I/R group rat heart nucleus albumen P65, the rat heart nucleus albumen P65 of each pre-administration group of CP is not significant yet to be changed.
Figure 13 A represents respectively to organize the variation of leukocyte adhesion factor CD11b, and sham operated rats leukocyte adhesion molecule CD11b average fluorescent strength is 31.11 ± 1.23.The leukocyte adhesion molecule CD11b average fluorescent strength of ischemia-reperfusion group rises to 41.86 ± 2.87, with the sham operated rats comparison and have statistical significance.The fluorescence intensity level that FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into back leukocyte adhesion factor CD11b in advance is respectively 40.66 ± 2.87,42.7 ± 3.9 and 35.07 ± 1.35.From as a result we as can be seen FUFANG DANSHEN DIWAN drop in advance the back CD11b expression there is no obvious influence.
Figure 13 B represents respectively to organize the variation of leukocyte adhesion factor CD18, and sham operated rats leukocyte adhesion molecule CD18 average fluorescent strength is 49.1 ± 3.59.The leukocyte adhesion molecule CD18 average fluorescent strength of ischemia-reperfusion group rises to 68.93 ± 4.21, with the sham operated rats comparison and have statistical significance.The fluorescence intensity level that FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into back leukocyte adhesion factor CD18 in advance is respectively 74.38 ± 4.83,73.59 ± 4.93 and 57.99 ± 3.8.From as a result we as can be seen FUFANG DANSHEN DIWAN 0.8g/kg can obviously suppress the expression of the leukocyte surface adhesion molecule CD18 that ischemia-reperfusion causes.
What Figure 14 represented is that Sham group, I/R group, CP0.1+I/R group, the CP0.4+I/R that laser-Doppler records organizes, the image of CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have obvious variation in this process.I/R group rat blood flow after pouring into again obviously reduces.CP0.1+I/R group rat heart blood flow is compared to some extent with I/R when pouring into 30min with 60min again and is recovered.CP0.4+I/R group, CP0.8+I/R group rat cardiac blood flow after pouring into again obviously recover.
The continuous variation of the expression Sham group of Figure 15, I/R group, CP0.1+I/R, CP0.4+I/R group, CP0.8+I/R group rat heart blood flow.Sham group rat heart blood flow does not have significant change in this observation process.I/R group rat heart blood flow is in ischemia and reduction significantly between flush phase again, and the CP0.1g/kg successive administration does not suppress the reduction of the rat heart blood flow behind the ischemia-reperfusion significantly.The reduction that can significantly suppress to pour into again the blood flow that causes in 60 minutes was being poured into 5 fens and poured into to the CP0.4+I/R group again.CP0.8+I/R group from pour into again 5 assign to pour into again 60 minutes during in, all can significantly suppress to pour into again the reduction of the blood flow that causes.
Figure 16 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart TTC colored graph when CP0.8+I/R group rat is poured into 60 minutes again.Sham group rat heart is not seen infarcted region, the visible bigger painted infarcted region that is white in color of I/R group rat heart.Reduce significantly in CP0.1+I/R group, CP0.4+I/R group and CP0.8+I/R group myocardial infarction zone.
Figure 17 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, heart myocardial infarction area statistic analysis result when CP0.8+I/R group rat is poured into 60 minutes again.I/R group rat heart muscle infarcted region area is 15.55 ± 2.14%.CP0.1+I/R group, CP0.4+I/R group and CP0.8+I/R group rat heart muscle infarct size be respectively 8.32 ± 1.42%, 7.65 ± 1.66%, 7.56 ± 2.57%, reduced the area of the myocardial infarction that ischemia-reperfusion causes significantly.
Figure 18 pours into 90 o'clock painted images of heart TUNEL again for Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat.The male myocardial cell of the Sham group accidental TUNEL of rat, I/R group rat heart can be observed the male myocardial cell of a large amount of TUNEL, and the TUNEL positive cardiomyocytes of CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group all reduces significantly.
Figure 19 represents Sham group, I/R group, CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat heart TUNEL positive cardiomyocytes result.Sham group rat TUNEL positive cardiomyocytes only is 0.36 ± 0.11/visual field, and I/R group rat TUNEL positive cardiomyocytes is 14.7 ± 1.17/visual field, significantly organizes more than Sham.CP0.1+I/R group, CP0.4+I/R group, CP0.8+I/R group rat TUNEL positive cardiomyocytes quantity are respectively 7.1 ± 0.54,7.23 ± 0.66 and 6.4 ± 0.44 (individual/visual field), all are markedly inferior to the I/R group.
Figure 20 A represents respectively to organize the variation of peripheral blood GA molecule CD11b.Sham operated rats peripheral blood GA molecule CD11b average fluorescent strength is 31.11 ± 1.23.Ischemia-reperfusion group peripheral blood GA molecule CD11bCD11b average fluorescent strength rises to 41.86 ± 2.87, is higher than sham operated rats significantly.FUFANG DANSHEN DIWAN 0.1g/kg, 0.4g/kg and 0.8g/kg drop into the back does not in advance have significant inhibitory effect to the increase of the expression of GA factor CD11b.
Figure 20 B represents respectively to organize the variation of leukocyte adhesion factor CD18, and sham operated rats leukocyte adhesion molecule CD18 average fluorescent strength is 49.1 ± 3.59.The leukocyte adhesion molecule CD18 average fluorescent strength of ischemia-reperfusion group rises to 68.93 ± 4.21, is higher than sham operated rats significantly.FUFANG DANSHEN DIWAN 0.1g/kg, be respectively 54.15 ± 2.09 0.4g/kg drop into the fluorescence intensity level of back leukocyte adhesion factor CD18 in advance with 0.8g/kg, 57.42 ± 2.16 and 56.73 ± 2.79, suppressed the expression of the leukocyte surface adhesion molecule CD18 that ischemia-reperfusion causes significantly.
The specific embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Drop pill of the present invention is that following method is made: Radix Salviae Miltiorrhizae extract 4-60 weight portion, Radix Notoginseng extract 2-18 weight portion, Borneolum Syntheticum 1-10 weight portion, press recipe quantity with Radix Salviae Miltiorrhizae extract, Radix Notoginseng extract and Borneolum Syntheticum fine powder mix homogeneously are stand-by, Radix Salviae Miltiorrhizae extract 4-60 weight portion, Radix Notoginseng extract 2-18 weight portion, Borneolum Syntheticum 1-10 weight portion is pressed recipe quantity with Radix Salviae Miltiorrhizae extract, and Radix Notoginseng extract and Borneolum Syntheticum fine powder mix homogeneously are stand-by, press recipe quantity, with Radix Salviae Miltiorrhizae extract, Radix Notoginseng extract and Borneolum Syntheticum are pulverized, and mix homogeneously is stand-by; Press medicated powder: substrate 1: 1-2 with substrate PEG6000 or PEG400 fusion after, it is even to add medicament mixed, 60-90 ℃ of insulation dripped/minute added in the methyl-silicone oil with 20-70, collects drop pill promptly.Preparation process advanced person of the present invention, stable and controllable for quality, determined curative effect.
Embodiment 2
The step of preparation process of FUFANG DANSHEN DIWAN of the present invention is: (1) is with Radix Salviae Miltiorrhizae and panax mixed or make water extract or alcohol extract separately; (2) described extracting solution being carried out predefecation handles; (3) further described extracting solution is carried out hyperfiltration treatment; (4) ultrafiltrate is concentrated, add Borneolum Syntheticum, make drop pill.
Claims (10)
1, FUFANG DANSHEN DIWAN treats and/or prevents application in the medicine of cardiac microcirculation disturbance and myocardial damage in preparation.
2, the application of claim 1 is characterized in that, described FUFANG DANSHEN DIWAN Main Ingredients and Appearance is the compound Chinese medicinal preparation that Radix Salviae Miltiorrhizae, Radix Notoginseng and Borneolum Syntheticum are formed.
3, the application of claim 1 is characterized in that, described application is that the disposable pre-input of FUFANG DANSHEN DIWAN can be improved ischemia-reperfusion microcirculation disturbance and myocardial damage.
4, the application of claim 1 is characterized in that, the described application cardiac microcirculation disturbance that to be the repeatedly pre-medicine of FUFANG DANSHEN DIWAN cause ischemia-reperfusion and the protective action of myocardial damage.
5, the application of claim 3 is characterized in that, described application is the cardiac microcirculation disturbance that the prevention ischemia-reperfusion causes, comprise the contraction that suppresses the coronary vasodilator arteriole, blood flow rate reduces, and albumin spills increase, the reduction of blood flow reduction and myocardial flow.
6, the application of claim 3 is characterized in that, described application is to improve ischemia-reperfusion microcirculation disturbance and myocardial damage, the expression of this effect and its inhibition GA molecule CD18, suppress the activation of MPO, the consideration convey of I-κ B-a degraded and P50 moves, and suppresses the myocardial cell accent and dies relevant.
7, the application of claim 1 is characterized in that, described application is the reduction that the muptiple-use pre-administration of FUFANG DANSHEN DIWAN can dosage prevents the cardiac blood flow that ischemia-reperfusion causes interdependently, suppresses the damage of myocardial cell.
8, the application of claim 4 is characterized in that, the repeatedly pre-administration of the FUFANG DANSHEN DIWAN that described repeatedly pre-administration is 0.1g/kg, 0.4g/kg and 0.8g/kg consumption can dosage suppresses the reduction of cardiac blood flow behind the ischemia-reperfusion interdependently.
9, the application of claim 4 is characterized in that, the repeatedly pre-administration of the FUFANG DANSHEN DIWAN that described repeatedly pre-administration is 0.1g/kg, 0.4g/kg and 0.8g/kg consumption can dosage suppresses the myocardial cell number of heart infarct size and apoptosis interdependently.
10, the application of claim 4 is characterized in that, the repeatedly pre-administration of the FUFANG DANSHEN DIWAN that described repeatedly pre-administration is 0.1g/kg, 0.4g/kg and 0.8g/kg consumption can dosage suppresses the expression of peripheral blood granulocyte adhesion molecule CD18 interdependently.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048818A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力制药股份有限公司 | Total salvianolic acid, panax notoginseng saponins, and compatibility thereof in prevention and treatment of diseases caused by microcirculatory disturbance |
| EP2586452A4 (en) * | 2010-06-28 | 2014-01-15 | Tasly Pharmaceutical Group Co | Use of a chinese medicine composition in preparing medicaments for treating secondary prevention of myocardial infarction |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048818A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力制药股份有限公司 | Total salvianolic acid, panax notoginseng saponins, and compatibility thereof in prevention and treatment of diseases caused by microcirculatory disturbance |
| EP2586452A4 (en) * | 2010-06-28 | 2014-01-15 | Tasly Pharmaceutical Group Co | Use of a chinese medicine composition in preparing medicaments for treating secondary prevention of myocardial infarction |
| US9211310B2 (en) | 2010-06-28 | 2015-12-15 | Tasly Pharmaceutical Group Co., Ltd | Use of a Chinese medicine composition in preparing medicaments for treating secondary prevention of myocardial infarction |
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